NZ236966A - 7-substituted androstan-3,17-dione derivatives and medicaments - Google Patents

Description

New Zealand Paient Spedficaiion for Paient Number £36966 \ A 2 3 6 £ Cc- ■ < 'fcs 1 H « 2 5 NOV 1993 NEW ZEALAND PATENTS ACT, 1953 No.-Date: COMPLETE SPECIFICATION AROMATASE INHIBITORS 3/ <\<\ //99;^ -P7We, ENDORECHERCHE INC., a corporation of Canada, of 2989 de la Promenade Ste-Foy, Quebec G1W 2J6, Canada hereby declare the invention for which we pray that a patent may be granted to p=r67us, and the method by which it is to be performed, to be particularly described in and by the following statement: - (followed by page la) 2 3 6 9 6 iQ AROMAIASE INHIBITORS BACKGROUND OF THE INVENTION This invention relates to novel steroidal aromatase inhibitors. More particularly, certain preferred embodiments of the invention relate to 7a-substituted androstenedione and androstanedione analogs which inhibit this enzyme.

Brief description of the prior art During the treatment of certain estrogen-dependent diseases, it is important to greatly reduce or, if possible, eliminate estrogen-induced effects. Alternative or concurrent therapy to administration of antiestrogens could involve attempts to block the production of estrogen such that none is available to activate receptor sites. The blockade of arcmatase, an enzyme that synthesizes estrogens from androgens (e.g. estradiol from testosterone) has been intensively studied in view of developing pharmaceutical drugs useful in the therapy of estrogen-dependent diseases, particularly breast cancer (Walsh, C., 1984, Ann. Rev. Biochem., 53, 493-535; Brodie, A.M.H. et al., 1977, Endocrinology, 100, 1684-1685; Covey D.F. et al. , 1981, J. Biol. Chera. , 256: 1076-1079; Covey D.F. and Hood, w.F. , 1981, Endocrinology, 108: 157-1599; Brueggemeier and Katlic, 1987, Cancer. Res. 47: 4548-4551; Henderson, 1987. J. Steroid Biochem. 27: 905-914; Humazawa et al., 1987. J. Steroid Biochem. 28: 337-344; Kruter et al., 1987, J. Steroid Biochem. 28: 139-145; Snider, C.E. and Brueggemeier, ^R.W., 1985, J. Biol. Chem., 262: 8685-8687; Sherwin, P.F. et al., 1989, 2 3 5 9 i • 2 J. Med. Chem. , 32, 551-658; Giudici, D. et al., 1988, J. Steroid Biochem., 30, these drugs have shown an "in vivo" inhibitory effect on the growth of mammary tumors (Spinola, P.G. et al., 1988, Breast Cancer Res. Treat., 12, 287-296; Brodie, A.M.H. et al., 1982, Adv. Exp. Med. 3iol., 138, 179-196; Brooks, S.C. et al., 1987, Cancer Research, 47, 4623-4629; Schieweck, K., 1988, Cancer Research, 48, 834-838); Schiavo et al., 1988, rund, Appl. Toxicol, 10, 329-334) but few of these have been used in women for the therapy of breast cancer. Examples of those which have been used are 4-hydroxy-4-androstenedione and aminogluthetimide (Coombes, R.C. et al. , 1984, The Lancet, 1237-1239; Brodie, A.M.H., 1987, J. Steroid Biochem., 27, 899-893; Goss et al., Cancer Res. 46: 4823-4826.; Brodie, A.M.H. and Santen, R.J., 1986, in Davies, S., CRC critical reviews in oncology/hematolo-gy, vol. 5, Boca Raton: CRC Press: 361).

Schweikert et al., 1986, US Patent N° 4,596,796, disclose the use of aromatase inhibitors for prophylaxis and/or treatment of benign prostatic hyperplasia.

U.S. Patent Ho. 4,822,528 discloses 4-substituted 6-alkylidenan-drostene-3,17-dione derivatives as aromatase inhibitors.

U.S. Patent No. 4,824,830 discloses 6- or 7- methyl-androsta-1 ,-4-diene-3,17-diones as aromatase inhibitors U.S. Patent No. 3,766,213 discloses the synthesis of retro-steroids and more particularly a method for the formation of the A-ring of retrosteroids.

Derivatives of 6-methylene-4-anarostene-3-ones are described by Petrow et al., 1983 (J. Steroid Biochem. 19: 1491-1502). > V* £ 0 fc ^ •l J Grunweil et al., 1976, Sreroids, 27, 759-771 and Solo, A.J. et al., 1982, Steroids, 40, 603-514 disclose the synthesis of a series of 7a-alkyltestosterone derivatives and describe their biological activities.

Brueggemeier, R.W. et al., 1978, J. Med. Chem. 21, 1007-1011 and Snider, C.E., Brueggemeier, R.W.,1985, J. Biol. Chem., 262, 8685-8687 disclose the synthesis and aromatase inhibitory activity of 7a-(4'-amino)phenyIthio-androstenedione derivatives.

Brueggemeier, R.W. and Li, P.K., 1988, Cancer Research 48, 6808-6810, disclose that 7a-(41-amino)phenylthio-4-anarostene-3,17-dione inhibits the growth of DMBA-induced mammary carcinoma in rats.

J.K. Davies et al. . Proc. Am. Ass. Cancer Res. 30, abs 1204, 1989, disclose the use of 4-'nydroxy-androstenedione in human.

K. Schieweck et al.. Proc. Am. Ass. Cancer Res. 30, abs 1205, 1989, disclose the use of CGS 16949A as anti-tumor agent.

R.W. Brueggemier et al.. Proc. Am. Ass. Cancer Res. 30, abs 1206, 1989, disclose the synthesis and activity of 7-alkyl and -aryl substituted derivatives of 4, 6-androstadiene -3, 17-diones and 1,4,6-androstatriene -3 ,17-diones.

R. De Coster et al.. Proc. Am. Ass. Cancer Res. 30, abs. 1207, 1989, disclose the use of novel triazole derivative R76713 as aromatase inhibitor.

U. Nickisch and H. Laurent in German patent D.E. 3612632 kl, 1987 disclose the synthesis of 7a-propylsteroids. 236966 J.A. Zderic in US parent "°3,485,828, 1569, disclose the synthesis of 6,7-ethylene and 6,7-substituted ethylene steroid derivatives.

New aromatase inhibitors capable of effectually blocking the activity of aromatase while minimizing side effects and maxiiaizing in vivo stability are desirable in the art for possible use in treating estrogen-sensitive diseases.

Objects of the invention It is an object of the present invention to provide a steroidal inhibitor of aromatase for therapeutic use, and pharmaceutical compositions thereof.

It is another object of the invention to provide methods of inhibiting aromatase using pharmaceutical compositions with good in vivo stability and/or low tendency to induce undesirable side effects. 2 3 6 9 6 Summary of i-he» invention The above and other objects are accomplished by providing aromatase-inhibiting compounds, and pharmaceutical compositions comprising therapeutically effective amounts of at least one of said compounds, wherein the aromatase inhibiting compounds have the following molecular structure. at the 4- and 6- positions, and wherein the dotted lines represent optional double bonds (4-androstene species being preferred); vherein R7 is selected frcm the group consisting of alkenyl, alkynyl, alkyl, epoxyalkyl, cyclopropyl alkyl and their halogeno-substituted derivatives.

All aromatase-inhibiting compounds herein have an anrirostenedione nucleus, the atom numbers and ring letters of which are as set forth above. The following conventions apply to structural formulae set forth herein. Unless specifically designated to the contrary, substi-tuents may have either a or p stereochemistry or, where valence permits may represent one substituent in a position and another in P position. Presence of optional double bonds are independent of each other. All structures include salts thereof. 2 7 £ /N fs o o ^ o 6 6 Atoms of any androstenedione nucleus for vnich no substituent is snovn or described nay optionally be substituted or uasubstituted. Those atoms having a defined substituent may optionally be further substituted by other substituents where their valence permits such further substitution. As used herein, the term "lover", when describing a chemicaJ. moiety means a moiety having 8 or fewer atoms. For instance, a "lower alkyl" a C. to C, alkyl. Any moiety of more than two atoms may be straight- or branched-chain unless otherwise specified.

A substitution at the 16 position is preferred in order to avoid the reduction of the 17-keto group which could cause the compounds to possess androgenic activity and potentially increase undesirable side effects in women.

R7 is preferably a substituent of fewer than 8 atoms, or even a shorter (Cj-CJ alkenyL (Cj-Cj alkynyl, (C1-CJ alkyl oar (C1-Ck) epaxyalkyl, cyclopropyl alkyl or halogeno—substituted derivatives thereof. In certain embodiments, R7 is in the alpna position and is CH^Y wherein Y is selected from the group consisting of —CH"CH2, —CECH, -CH°CCHj, -CEC-CH. , -CH-CH and -CH-CH.. In some embodiments, there are a \/ \i CE, 0 plurality of points of unsaturation in the C7 substituent.

The aromatase inhibition is preferably a 4-androstenedione substituted as taught herein. Other embodiments include but are not limited to 5cc—androstanedione and Sp—androstanedione. The aromatase inhibitors 236 9 6 6 7 of the invention nay be used to treat estrogen-sensitive diseases by, for example, inhibiting the production of estrogen. These diseases include, but are not limited to breast cancer, ovarian cancer, endometriosis, benign breast disease, gynecomastia, uterine fibroma, precocious puberty and benign prostatic hyperplasia. The pharmaceutical compositions may also be useful in the control of fertility in women and infertility in men.

Aromatase-inhibiting compounds of the invention may also inhibit the activity of other estrogen-producing enzymes, especially, but not limited to 17p-hydroxysteroid dehydrogenase and estrogen sulfatase.

In one alternative embodiment, the 16 carbon in the D ring is replaced with heteroatora capable of inhibiting reduction of 17-keto. /» 3 Detailed description of certain preferred embodiments In certain preferred embodiments of the invention, therapeutic compositions may comprise cne or core compounds represented by the formula II: Wherein R7 is selected from the group consisting of alkenyl, alkynyl, derivatives and wherein B is a stabilizing substituent which inhibits reduction of the 17-keto group. B is preferably a poor leaving group and is preferably even capable of steric hindrance of 17-keto reduction, (either by decreasing the arosatase-inhibiting compounds affinity for 17p-hydroxysteroia dehydrogenase, or otherwise) In preferred embodiments, B is selected from the group consisting of: methyl, ethyl, iso-propyl, t-butyl, phenyl, tolyl and trip'nenylmethyl.

Alternatively, reduction of 17-keto may be inhibited by placing an appropriate heteroatom at the 16 position of the D ring.

By way of example, some preferred aromatase inhibitors in accordance with the invention include but are not limited to are: cyclopropyl alkyl, epoxyalkyl and their halogeno-substituted 2 * £ 0 £ ~ V vi 0 9 7a-allyl-4-androsten-3,17-dione 7p-allyl-5p-androstan-3,17-dione H 7a-allyl-5a-androstan-3,17-dione 7cc-allyl-16-oxo-4-androstan-3 ,17-dione 7a-allyl-16-oxo-5p-androstan-3,17-dione 23 6 9 .& & 7a-aliyI-i5-oxo-5a-androstan-3,17-dione 7a- (2-propynyl) -4-anarosten-3 ,17-diorxe 7a- (2-propynyl) -5-a-andros tan-3 ,17-dione 7a- (2-propynyl) -16-oxo-4-androstan-3^ 17-dione c. i - 7 OCT i??3" ' 236966 li When aromatase inhibitors in accordance with the invention are used in the treatment of estrogen-related diseases, they are preferably administered at a dosage from about 5 rag to about 2000 mg of active expedient (i.e. aromatase inhibitor) , per day per 50 kg of body weight, most preferably from about 1.0 rag to about 20 mg per day per kg of body weight.

The aromatase inhibitors are preferably prepared as pharmaceutical compositions together with pnarmaceutically acceptable carriers and diluents. When prepared for parenteral injection, an inhibitor of aromatase activity may be prepared in a carrier preferably selected from the group consisting of saline, water, aqueous ethanol and oil.

When a pharmaceutical composition of the invention is prepared for oral ingestion, the composition preferably includes at least one inhibitor of aromatase activity wherein the total concentration of all such inhibitors in said pharmaceutical composition is from about 1% to about 95% of the composition (by weight), and preferably from about 5% to about 20%. The composition preferably further includes a pharmaceu-tically acceptable diluent, for example, starch or lactose with or without tartrazine. Slow release pharmaceutical products comprising the novel inhibitors of aromatase activity may be incorporated into slow release pharmaceutical products which, other than addition of the novel inhibitors, may be prepared by known methods and administered orally as well as parenterally.

In certain alternative embodiments, the pharmaceutical composition of the invention may be formulated for sustained release in accordance with known techniques. These sustained release formulations are preferably prepared in an appropriate manner for either oral, intramuscular, or subcutaneous administration. 2 3 6 9 12 Set forth below is a detailed description of preferred synthetic techniques for producing certain preferred aromatase inhibitors in accordance with the invention. mHPT.KS OF SYNTHESIS OF PREFERRED AROMATASE INHIBITORS (SEE SCHEMA I AND SCHEMA II) Instrumentation The IR spectra were taken on a Perkin-Elmer 1310 spectrophotometer. Proton NMR spectra were recorded on a Varian EM-360A (60 MHz) or a Varian XL-200 (MHz) instrument. The following abbreviations have been used: s, singlet; d, doublet; dd, doublet of doublet; t, triplet; q, quadruplet; and m, raultiplet. Chemical shifts are reported in <1 values relative to tetraraethysilane (TMS) or chloroform as internal standard.

For flash chromatography, Merck-Kiesel gel 60(230-400 mesh A.S.T.M.) was used. All solvents used for chromatography have been distilled. Unless otherwise indicated, starting material and reagents were obtained commercially and were used as such or purified by standard means. 7a-allyl-17{3-hydroxy—4-androsten-3-cme, "EM 172".

To a solution of 17p-acetoxy-4,6-androstadien-3-one 1 (2.0 g, 6.1 mraol) in dry dichlororaethane (130 ml) was added titanium tetrachloride (5.36 g, 3.1 ml, 28 mmol) at -70°C. The reaction mixture was allowed to stir for 5 rain. Then, a solution of allyltriraethylsilane (4.0 g, 5.6 ml, 35 ramol) in dry dichlororaethane (10 ml) was added pver a period of 10 rain and the resulting solution stirred during 1 hour at 2 3 6 9 13 -70°C followed by 1 hour at -20°C. Afterwards, the solution was diluted with 150 ml of ether and was washed with water (6 x 100 ml) , dried, filtrated and concentrated to yield the crude 17p-acetoxy-7a- allyl-4-androsten-3-one which was immediately hyarolyzed.

To a solution of crude acetate in methanol (100 ml) was added aqueous hydrochloric acid (10 ml, 5N). The reaction mixture was heated at reflux for 1 hour. Then, most of the methanol was evaporated and the residue was transferred into a separatory funnel with a mixture of ether: dichloromethane (100 ml, 1:1) and water (50 ml) and was then washed thoroughly with water. The organic phase was dried, filtered and concentrated to a solid. The crude material was purified by flash chromatography (nexane: acetone, 9:1 and 4:1) to yield 630 mg, 32% of compound EM 172. m.d. 208-210°C, IR (KBr) \> cm"1: 3550-3250 r • max (OH), 1645 (C=0), 1610 (C=C); lH-NMR (fippra): 5.72 (lH,s,-CH=C-) , 5.65 (lH,m,-CH=CHj), 5.00 (2H,ra,-CH=CH.), 3.67 (lH,t,J= 8.5 Hz,-CH0H), 1.22 (3H, s,-19CH3), 0.81 (3H.s,-iaCH,). 7a-allyl-4-androsten-3,17-dione, "EM 173" A mixture of EM 172 (100 mg, 0.3 mmol) , pyridinium chlorochroniate (100 mg, 0.46 mmol), sodium acetate (75 rag, 0.91 mmol) ana 4A molecular sieve (200 rag) in dry dichlororaethane (5 ml) was stirred at 25°C for 2 hours. Then, the reaction mixture was filtered through a silica gel pad with ether as eluent and the filtrate was evaporated to a solid. The residue was recristalized from dichloromethane and ether to 2 3 5 9 6 14 I give 81 rag, 31% of the desired diketone EM 173. ra.p. 2i9-221°C, IR (KBr) u era*1: 1720 (C=0, ketone), 1670 (C=0, enone) , 1635 (C=C) , max 1607 (C=C); iH-NHR (6ppm): 5.74 (lH,s ,-CH=C-), 5.68 (1H,m,-CH=CH3), 5.03 (2H, ra,-CH=CH,), 1.23 (3H, s, -,9CH,), 0.93 (3H, s,-,aCH3). 7 a—allyl-5p-androstan-3,17-dione, "EM 176" and 7a-allyl-17p-hydrcnEy^-5p-androstan-3-one, "EM 178".

To a stirred suspension of copper iodide (57 mg, 0.3 nmol) in dry tetra'nydrofuran (5 ral) cooled to 0°C was added nethyllithiun (215 ^il of a 1.4 M solution in ether, 0.3 ramol). The reaction mixture was cooled to -50°C and then 2 ral of hexamethylphospnoramide and diisobu-tylaluminura hybride (4 ml of a 1M solution in hexane, 4 mmol) were added successively. The mixture was stirred for 30 min. at -50°C and EM 173 (50 rag, 0.15 ramol) in 2 ml of tetrahydrofuran was added. After being stirred at -50°C for 2 hours, the solution was diluted with 30 ral of ether and was washed with aqueous hydrochloric acid (2 x 5 ral, 10% aqueous) and water (6 x 15 ml). The ethereal phase was dried, filtrated and concentrated to an oil. The residue was purified by column chromatography (hexane: acetone, 9:1) to give EM 176, 11 mg, 22%; EM 173, 8.5 mg, 17%; EM 178, 2.5 mg, 5% and EM 172, 13.5 mg, 26% obtained in that order from the column. N.B. EM 176: IR (neat) v era'1: 1730 (C=0) , 1710 (C=0) , 1632 (C=C) ; lH-NMR (ippra) : 5.74 raax (lH,ra,-CH=CH,) , 5.04 (2H, m, -CH=CH2), 2.68 (1H, d of d, J = 13.20 Hz 23 6 9 I , and J= 15.55 Hz, 0 = C-CH H CH-, 1.11 (3K,s,,CH,), 0.90 (3H, —ax eq I! s,-..CH,). EM 178: IR (neat) v era" 1: 3600-3150 (OH), 1708 (C=0) , -» -J max 1635 (C=C); lH-NMR (6ppm): 5.72 (lH,ra,-CH=CKJ , 5.03 (2H,mt-CH=CHa) , 3.68 (lH,t, J= 8.40 Hz, -CHOH), 2.70 (1H, dd, J= 13.55 Hz and J= 15.55 Hz,0=C-CHaxHe CH-), 1.10 (3H,s1,CH3) , 0.79 (3H.S,-j,CH3). 17a-allyl- 17^- (t-butyldimethylsilyloxy)-4-androsten-3-one, "2" To a solution of EM 172 (420 mg, 1.28 mmol) in a mixture of dry tetrahydrofuran and dimethylformamide (14 mi, 1:1) cooled at 0°C was added imidazole (700 mg, 10.2 mmol) and t-butyldimethylsilyl chloride (770 mg, 5.1 mmol). The reaction mixture was allowed to warm up to 25°C and was stirred during 20 hours. Then, the mixture was diluted with 150 ral of ether and was washed thoroughly with water. The ethereal phase was dried, filtrated and concentrated to an oil which was purified by flash chromatography (hexane: acetone, 9:1) to yield 546 mg, 96% of silylether 2. IR (neat) v max cm" 1: 1670 (C=0) , 1632 (C=C) , 1610 (C=C) ; iH-NMR (<5ppm): 5.70 (lH.s ,-CH=C-) , 5.67 (1H,ra,-CH=CH3), 4.99 (2H,m,-CH=CH,) , 3.56 (lH,t,J= 8.5 Hz,-CH0Si-), 1.19 (3H,s,-l9CH3), 0.87 (9H,s,-C(CH3)3) , 0.75 (3H,s,-1#CH3) , 0.00 (6H.s,-Si(CH,),). > 2 3 6 9 16 17p-hydroxy-7a-propy 1-5a-androstan-3-one, "EH 185" A large excess of lithium wire (405 rag, 58 mmol) cur into short sections was added to 12 mi of freshly distilled ethylenediamine cooled in a water bath to 15°C. The mixture was stirred until dark blue (5 min.). Then, a solution of enone 2 (150 rag, 0.34 mmol) and t-butyl alcohol (435 p.1) in 2.5 ral of dioxane was added aropwise rapidly; additional dioxane was used to complete the transfer. The reaction mixture was stirred for 45 rain, while being maintained below room temperature by addition of ice to the water bath. Afterwards, ammonium chloride (1.25 g) and water (40 ml) were added successively to quench the reaction. The resulting mixture was extracted with ether (3 x 30 ml) . The ethereal phase was washed thoroughly with water, dried, filtered and concentrated to an oil which was directly hydrolyzed.

To a solution of crude silyl ether in methanol (5 ml) was added aqueous hydrochloric acid (2 ral, 10% aqeous). The solution was heated at reflux for 1 hour. Then, the reaction mixture transferred into a separatory funnel with ether (50 ral) and water (20 ral) was washed with water (6 x 20 ral) . The organic phase was dried, filtrated and concentrated to a solid. The crude material was purified by flash chromatography (hexane: acetone, 9:1) to yield 38.5 rag, 34% of EM 185. m.p. 154-156°C, IR (KBr) y cm"1: 3560-3100 (OH), 1700 (C=0) ; max i'H-NMR (6ppra): 3.65 (lH,t,J= 8.1 Hz,-CHOH) , 1.04 (3H,s,CH3) , 0.88 (3H,t,J= 6.7 Hz, -CH2CH3), 0.76 (3H,s,-l8CH3). 2 3 6 9 6 17 7a-allyl-17p-hydroxy-5a-androstan-3-one, "EM 197" A solution of EM 172 (500 mg, 1.52 mmol) 2nd t-butyl alcohol (5 ml) in dioxane (25 ml) was added dropwise with stirring to a solution of lithium (68 mg, 9.7 mmol) in liquid ammonia (100 ml) over a period of 2 minutes. The reaction mixture was allowed to stir for another 5 minutes. The lithium amide formed was neutralized by the addition of ammonium chloride (2 g) and the ammonia was allowed to evaporate. The residue was dissolved in ether (200 ral), washed with water (6 x 50 ml), dried, filtrated and concentrated to an oil. The residue was purified by flash chromatography (hexane: acetone, 4:1) to yield 254 mg, 50.5% of hydroxyketone EM 197: m.p. 170-172°C, IR (KBr) \j cm'1 : 3560-3 300 (OH), 1695 (C=0) , 1633 (C=C) ; 1H-NMR (flppm) : max .66 (lH.m.-CH-CHj) , 4.96 (2H,m,-CH=CH3) , 3.65 (lH,t,J= 8.30 Hz,-CHOH), 1.04 (3H,s,-l9CH,) , 0.77 (3H,s,CH,) . 7a-sllyl-5a—siidrostan-3,17-dione, "EM 198" The preparation of this diketone (EM 198) was performed as described for diketone EM 173 (vide supra) with the following quantities: EM 197 (56 mg, 0.168 mmol), pyridinium c'nlorocnromate (110 rag, 0.5 ramol), sodium acetate (83 mg, 1 mmol) and 4A molecular sieve (110 mg) , dichlororaethane (5 ml), 0°C - 25°C, 4 hours. The 2 3 6 9 18 residue was purified by flash chromatography (hexane: acetone, S:l) to give 35 rag, 71% of diketone EM 158. IR (neat) \j riav cm-1: 1732 (C=0), 1709 (C=0) , 1635 (C=C); lH-NMR (fippm): 5.69 (lHfm,-CH=CH.) , 5.00 (2H,m,-CH=CH.), 1.06 (3H,s,-l9CH,), 0.89 (3K,s,-j.CH,). 7a-allyl-3,3-ethylenedio3cy-17p-hydrQX5^-5-aiidrostane A mixture of EM 197 (620 rag, 1.87 mmol), ethylene glycol (200 mg, 180 ill, 3.22 maol) and p-toluenesulfonic (10 rag, 0.058 mmol) dissolved in 80 ml of dry benzene was refluxed (Dean-Stark) for 2 h 45 rain, under nitrogen. Then, ether (100 ml) was added and the resulting solution washed successively with sodium carbonate (2 x 30 ral, 5% aqueous) and with water (4 x 30 ml). The organic phase was dried, filtered and concentrated to a solid. The residue was purified by filtration on silica with hexane:acetone (4:1) and dichloromethane to give 550 mg, 78% of dioxolane. IR (KBr) ujnax cm"1: 3600-3100 (OH), 1100-1080 (C-O); iH-NMR (6 ppra): 5.69 (lH,m,-CH = CHa), 5.00 (2H,m, -CH = CHa), 3.92 (4H,s,-OCHjCHjO-), 3.64 (lH,t,J = 8.20 Hz,-CHOH), 0.84 (3H,s,-l9CH,) , 073 (3H,s,-l,CH3) ; MS m/e (70 eV): 374 (M*). 17f}-hydroxy—7a-(2,3'-epoxypropyl)—5a—androstan—3—one ("EM 206") A mixture of above olefin (380 mg, 1.02 ramol), ra-chloroperbenzoi-cacid (1.2 g, 50-60%, 3.47 raraol) and sodium acetate (3.8 g) in dry 2 3 6 9 is « > dichloromethane (30 ml) was stirred for 24 h at 25°C. Then, ether(100 ml) was added and the resulting solution washed successively with a solution of sodium carbonate (3x, 50 ml) and with water (3 x 50ml). The organic phase was dried, filtrated ana concentrated to a solid. The residue was purified by flash chromatography (hexane:acetone, 4:1) to give 115 mg, 29% of the eooxide as a solid. IR (KBr) v cm-1: • TT»a V 3550-3100 (OH), 1100-1080 (C-O); 1H"NMR (s ppm): 3.93 (4H,s,"0CH3-CH:-0-), 3.62 (1H, t, J - 8.25 Hz,-CHOH) , 2.90 (1H, massive, -CHOCH,-), 2.77 (1H, dt apparent, J = 16.2 Hz and J = 4.7 Hz, CHOCHH) 2.48 (1H, m, CHOCHH), 0.86 (3H,s,-l# CH,) , 0.74 (3H,s, -laCHs); MS ra/e (70 ev): 390 (MO.

This compound was hydrolyzed with p-toluenesulfonic acid in acetone and the 17[3-hydroxy group oxidized with Jone's reagent into the (2',31-epoxypropyl)-5a-androstan-3,17-dione ("EH 206").

SCHEME I 2 3 6 9 6 6 OAc S-/ EM 173 O EM 176 EM 172 OTBDMS H EM 198 EM 199 2 3 6 9 6 6 SCHEME II 7a-£ilyl-I6-oxo-4androst:an-3 ,17-dione ("EM 230") a) S0C1,, pyridine; b) CH3=C(0Ac)CH3, p-TsOH, A; c) 1) 0,, CH.C1,, AcOK, -70°C, 2) (CH,),S; d) CH,N3, Et.O; e) NaBHt, EtOH; f) Na.CO,; g) > Zn, AcOH; h) Jcne's reagent; i) dichloroaicyanobenzoquir.one, H*. ^ Dioxane; j) TiClt, allyltrimethylsilane, CH,C13, -70"C to -20°C.

SCHEME II 2 3 8 9 6 $ 22

Claims (4)

1. fy ' ■/ f. r\_ *- \j '0 'd 23 Efficacy of preferred aromatase inhibitors synthesized in accordance with examples: The compounds synthesized above have been tested by the assay described by Thompson and Siiteri (J. 3iol. Chem. 249, 5364- 5372, 1974) slightly modified as set forth below, and have been found to be effective inhibitors of the activity of placental aromatase. Microsomal aromatase was obtained from human placenta. A reaction vessel was prepared containing microsomes (0.125 mg protein/ml), NADPH (0.625 raM) , [JH]androstenedione (0.33 pM) and increasing concentrations of the potential inhibitor being tested in buffer (800 ul) (0.1M KHjP04, pH 7.5). The conversion of androsteneaione to estrone was allowed to proceed at 37°C for 30 min and stopped by addition of 200 pi of charcoal suspension (25 mg/ral) in the same buffer. After cen-trifugation, tritiated water contained in 100 ul of supernatant was counted. The amount of tritiated water is proportional to the amount of transformed [3H] androstenedione. Ki values were calculated by the Dixon method (Dixon, M., 1953, Biochem. J. 55, 170-171) and reported "* *"* 1 belowt Potency of selected compounds as inhibitors of aromatase activity TABLE I Compound Ki (uM) EM 173 EM 176 EM 198 0.2 0.9 0.6 As shown in Table 1, each of EM 173, EM 176 and EM 198 showed aromatase inhibitory activity. WHAT/tfWE CLAIM IS Wthifc j q nl i irae.il La 2>; 6 9 6 8 1) An aromatase-inhibiting compound represented by the following » molecular structure: O n 13 16 19 2 15 wherein each dotted line independently represents an optional double bond; wherein R7 is selected from the group consisting of alkenyl and alkynyl.
2. 2) A pharmaceutical composition comprising a phamaceutically acceptable diluent or carrier and therapeutically effective amount of the aronatase-inhibiting compound of Clain 1.
3. 3) The compound of Claim 1 where the A/3 ring junction of said mole' cular structure is in trans configuration.
4. 4) A pharmaceutical composition comprising a ph2..uaceutically acceptable diluent or carrier and therapeutically effective amount of the aromatase-inhibiting compound of Claim 3. . > t > ' ' * • I > / V £ ft £ ^ 5) The compound of Claim 1 wherein -he A3-ring structure of said molecular strucutre is in cis configuration. o) A pharmaceutical composition comprising a pharmaceutical!/ acceptable diluent or carrier and therapeutically effective amount of the aromatase-inhibiting compound of Claim 5. 7) Tne compound 7a-allyl 4-anarosten-3,17-dione: "EM 173" 8) A pharmaceutical composition comprising a pharmaceutically acceptable diluent or carrier and therapeutically effective amount of the aromatase-inhibiting compound of Claim 7. 9) The compound 7a-allyl-5f5-androstan-3,17-dione: "EM 176" H 10) A' pharmaceutical composition comprising a p'narmaceutically acceptable diluent or carrier and therapeutically effective amount of the aromatase-inhibiting compound of Claim 9. S I ' I c:., r;3 11) The ccmpour.d 7c.-ailyl-5c-androstar.-3 , 17-dione: "EM 198" 12) A pharmaceutical composition comprising a pharmaceutical^ acceptable diluent or carrier and therapeutically effective amount of the arcmatase-irjiibiting compound of Claim 11. 13) The compound 7a-propyl-5c-androstan-3,17-dione: "EH 199" 14) A pharmaceutical composition comprising a p'narmaceutically acceptable diluent or carrier and therapeutically effective amount of the aromatase-inhibiting compound of Claim 13. 15) The compound of Clain 1 where all ate=s of R7 are separated froa a ring carbon of the 5 ring of said ccspound by no nore than seven intervening atens. 16) A pharmaceutical composition comprising a pharmaceutical^ acceptable diluent or carrier and a therapeutically effective enount of the aromatase-inhibiting compound of Claim 15. 69 6? 28 17) The compound of Claim 15 wherein R7 includes at; least two positions of unsaturaticn. 18) A pharmaceutical composition ccraprising a pharaaceutically acceptable diluent or carrier and therapeutically effective amount of the aromatase-inhibiting compound of Claim 17. 19) The compound of Claim 1 wherein R7 is in the alpha position and is _y wherein Y is selected from the grouo consisting of: 2 -CH-CH,, -CH-CHCHj and -C=C-CH3. 20) A pharmaceutical composition comprising a pharmaceutical ly acceptable diluent or carrier and a therapeutically effective amount of the aromatase-inhibiting compound of Claim 19. 21) An aromatase-inhibiting compound having the following molecular structure: wherein B is methyl or ethyl; wherein each dotted line independently represents an optional double bond; 29 23 5 Q 6 6 wherein R7 is selected from the group consisting of alkenyl and alkynyl. 22) A pharmaceutical cexposition comprising a pnaraaceutically acceptable diluent or carrier and therapeutically efrective amount of the aromatase-inhibiting compound of CLaim 21. 23) The compound 7a-(2-propynyl)-4-androsten— 3 ,17-dione: "EH—235" o 24) A pharmaceutical ccnposi~ion comprising a phamaceutically accept able diluent or carrier and therapeutically effective amount of the aromatase-inhibiting compound of Claim 23. 25) The compound of claim 1, wherein R7 is alkenyl. 26) The pharmaceutical composition of claim 2, wherein R7 is alkenyl. 27) The compound of claim 1, wherein R7 is C^-Q alkenyl. 28) The pharmaceutical composition of claim 2, wherein R7 is C^-C^ alkenyl. 29) The compound of claim 21, wherein R7 is alkenyl. r 30) The pharmaceutical composition of claim 22, wherein R7 is alkenyl. ~ ^ i I? 31) The compound of claim 21, wherein R7 is alkenyl. 32) The pharmaceutical composition of claim 22, wherein R7 is Q-Q alkenyl. o 9 6 & 30 33) The compound of claim 1, wherein R7 is C,-C4 alkenyl or C,-C4 alkynyl. 34) The pharmaceutical composition comprising a pharmaceutically acceptable diluent or carrier and a therapeutically effective amount of the aromatase inhibiting compound of claim 33. dated this HU day of CcAi^' c- i " . ' " M ,, I he 4 i C> / J "7 OCT [?93